Noroviruses (NVs) are the most important cause of non-bacterial epidemics of acute gastroenteritis, which is a common cause of morbidity and mortality worldwide. In US, outbreaks of NV-related gastroenteritis are estimated to affect 23 million people annually, representing a significant threat to public health. With the goal of developing novel therapeutic approaches for the treatment of NV infection, this proposal aims to identify small molecules that can effectively destabilize and/or disrupt the assembly of the NV capsid. Targeting NV capsid represents a novel strategy that has been successfully applied to other viruses in recent years. The proposed approach stems from our previous collaborative efforts and builds on expertise in both NV research and computational studies of proteins and their interactions. We propose an integrated strategy that uniquely combines: i) a novel computational framework for modeling and ranking of hot spots within NV capsid proteins; ii) virtual screening for small molecules targeting such sites; iii) experimental validation of top hit using newly established functional and structural assays and different model systems. Mapping of functional and structural hot spots will utilize the wealth of sequence and structural data on NVs. Ranking and assessment of the predicted hot spots will be further enhanced by integrating evolutionary, structural and functional information with modeling of conformational changes and allosteric effects associated with oligomerization of capsid proteins. Validation and further characterization of such identified hot spots will be performed by using mutagenesis, in vitro binding and in vivo functional assays. Virtual screening using state-of-art docking simulations, cheminformatic methods and comprehensive libraries of drug-like compounds will be used to identify candidate molecules targeting top ranking sites. Experimental validation of top hits will be performed using in vitro binding and in vivo functional assays developed by us recently for the assessment of NV capsid assembly. In order to facilitate this and future applications to other viruses, a versatile interface for automated analysis of structural and functional hot spots within capsid proteins will be integrated into our Polyview-MM portal.